Nitrated estrogenic hormones



Patented Mar. 30, 1948 UNITED;

NITRATEI J E STBQGENIQ HQEMQNE S. Joseph, B. Niederl, Brnoldyp,,N.Y

No Drawing. Application Aprill28, 219.45.,-

s same.590.5175;

4 Claims... (01.

1 This invention relates to new estrogenic com pounds and to methods ofvproducing the same. More. particularly it relates to'nitratedestrogeiiic hormones, to various derivatives thereof, and to methods ofproducingthe same from crude estrogenie extract deriVed from thecustomary urinarysources as well as from partially or"com 'oletelypurified individual phenolic hormones, phenolic-- hormone mixtures, orderivatives thereof.

Basically considered the instant'process isseen to involvetheintroduction of one or more nitro' groups into the benzene ring ofestrogenic her-- mones using especially developed micro and semimicroprocedures. Essentially the process in volves treatment of theestrogenic' hormone dis"- solved in an inert solvent, with one ortwo'mols' natural sources but also such compounds as estrone, estradiol.estriol, as well i as mixtures thereof.

The term estrogenic hormones alsoincludes the the ethers and esters ofthe phenolic estrogenic hormones. It also includes carboxylated or;halo1-" genated estrogenic hormones, with or without a free phenolichydroxy group.

Example I.Mono-mtro estrone from pure estrone One gram of pure estroneis dissolved in 25 ml. of boiling glacial acetic acid. This solution,while still hot but before crystallization sets in, is treated with onemol equivalent of nitric acid, as follows:

A micro dropping pipet is standardized first by counting the number ofdrops necessary for forming one cc. Pipets which give 40 drops for oneml. or drops for 0.25 ml. have been found most convenient. Three dropsof concentrated 70% nitric acid (spec. grav. 1.42) are added from amicro dropping pipet standardized as described above to the previouslyprepared hot glacial acetic acid solution of the estrone, After theaddition of the first three drops, the reaction mixture is thoroughlyshaken and allowed to cool further by immersing the reaction flask incold water. An additional 7 drops of the nitratingml.ohcoldglacialacetic acid, they aretransferr 2 agent are then slowlyadded followed by; a -thor ah mi insaficrihea dit on racles drop.

The reactionmixtureis then set aside at roorn temperature" to allow thecrystallization of the mono-nitro estrone. The crystals usually appearafter the reaction mixture has stood fonabfllltfi hours. The. crystals.are then separated by q-suc tionrfiltration,througha semi-microglassiritt 1g; funnel. After washingv of the orystals with a f d toaporoustil'e (3. 'x3'f and 1%! thick) They.- a e; coveredby a watjchglass and, are allowed; at room temperature; in all; When compl t eyeecrysta l zed-from boil n e I acetic acid; M. P.1'258 C. (uncor.).

Following the above semiemi lto, nltration pro cedure the're appearstobe nojoxidationwhich s; mostunusual with a compound of :SQcomplicated, a' structure; and the. yieldappears tobejneaul,

quantitative.

Applying. micro and. semi-micro technics (using sealed capillaries, orcentrifuge. cones). theimonw nitro'estror'i'eiis converted intovarious;ester s ;-such as 'the#acetate, propionate, butyrate, benzoate,

' nitro-be'nzo'ate, and 3','5'-.di nitro benzoate, as well.

, derivatives ofaboilizig glacial acetic. ac

as ethers such as methyl ether, the ethyl etheror; Z F-Q Q RZ th??- Themono-nitro estrone itself as well as its exhibits interestingphysiological detects, 1

Example II Di.-nz'tro cstrone from. pure est-pone e am orpure estrqnaisd ssq ed ll. 1 W i1e i h, ft 1i before crystallizat n sets,-

wenty. dres 1 m9! equivalentsxof concentrated nitr acid e. added in themanner and by means of the standardized micro dropping pipet asdescribed in Example I.

The reaction mixture is again allowed to stand until the di-nitroestrone has crystallized out, or the reaction mixture is poured into 250ml. of distilled water. The di-nitro estrone thus separates as aflocculent yellow precipitate, which is filtered off through a flutedfilter paper and washed thoroughly with distilled water. After thefilter paper with the precipitate has dried in air, the precipitate istransferred to a porous tile for completion of the drying process and isfinally recrystallized from hot glacial acetic acid, boiling methylalcohol, or other organic solvents. M. P. 275 C. (uncor.)

Using appropriate microor semi-micro procedures the di-nitro estrone isconverted into esters (acetate, propionate, benzoate, etc.) and ethers(methyl-, ethylor benzyl).

Example III.Monoand di-m'tro estradiol One gram of estradiol isdissolved in 20 ml. of boiling glacial acetic acid. The solution isallowed to cool until the first signs of crystallization begin toappear. At this moment dropwise addition of one mol of nitric aciddrops) is begun, utilizing the standardized micro dropping pipet, andfollowing the procedure as given in Example I.

After standing for 48 hours the reaction mixture is poured into 250 ml.of distilled water and M the precipitate of mono nitro estradiol isfiltered off, washed repeatedly with distilled water and" placed onporous tile. M. P. 175 C. (uncor.).

Substituting estriol for the estradiol produces the correspondingmonoand di-nitro estriol.

Example I V.--Monoand di-m'tro estrone'from technical estrone One gramof partially purified estrone is dissolved in ml. to ml. of boilingglacial acetic acid. This solution is then cooled and while still clearone or two mols equivalent of concentrated nitric acid are added in amanner similar to that described in Example I (10 or 20 drops ofconcentrated nitric acid, respectively, from the identicalmicro droppingpipet.)

The reaction mixture is then set aside at room temperature for 48 hours.After this time the reaction mixture is poured into 250 m1. of distilledwater whereby the nitrated estrone precipitates out as a fiocculentyellowish white precipitate. This precipitate is filtered 01f throughfluted filter paper, repeatedly washed with distilled water, placed onporous tile and allowed to dry at room temperature. The material at thisstage is a yellowish white powder, which may be further purified byre-crystallization from boiling methyl alcohol :or glacial acetic acid.7

Example V.-Nz'tration of estrogem'c hormone extract One gram ofsemi-solid black tar derived from customary urinary sources, whoseprincipal constituent is a mixture of the natural female hormones, isdissolved in 15 m1. of boiling glacial acetic acid. The solution isallowed to cool until the first signs of inhomogenuity are observed. Atthis moment 10 drops of concentrated nitric acid are added dropwise fromthe standardized 4 micro dropping pipet, as described in Example I.

The reaction mixture again is allowed to stand. for 48 hours and thenpoured into 250 ml. of distilled water. A brownish semi-solid precipi--tate is formed which may be further purified by fractionalcrystallization from methyl or ethylalcohol, benzene or chloroform; orthe crude mono-nitro estrogenie hormone mixture is used directly forfurther chemical transformation.-

, The semi-micro nitration process as described in the preceding 5examples is not limited to the types of hormones cited therein but isequally applicable to esters and ethers of the phenolic 'estrogenichormones, as well as to non-phenolic but benzenoid sterols. Unsaturatedcompounds,

however, should not be employed for nitration purposes.

In cases where ring or side chain unsaturated hormones or sterols are tobe subjected to nitra tion, it is desirable to first remove theunsatura-i tion by subjecting such compounds to hydrogena l tionprocedures prior to nitration.

The preceding nitration process is also ble tor benzenoid hormones orsterols possessing solvent, at an elevated temperature, tothe othergroups instead of or in addition to the. phenolic hydroxyl group, thuscarboxylatedror. halogenated compounds may be equally well subjected tothe foregoing nitration procedure. o

It will be understood that the examples disclosed herein areillustrative, and that the invention is not limited to the specificreactants and reaction conditions disclosed but extends to equivalentsencompassed within the scope of the invention which is defined by theappended claims. I

1. A process of producing a nitro estrone which comprises subjectingestrone'dissolvedin an inert action of concentrated nitric acid. r V r2. Nitrated estrone.

3. Mono-nitro estrone.

4. Di-nitro estrone. v

' r JOSEPH B. NIEDERL.

REFERENCES crTEn A The following references are of record'in' the fileof this patent: V

.UNITED STATES PATENTS 7,

Number Name Date 1,412,707 Richardson Apr. 11, 1922 2,207,727Galloway-1. July 16,19 1!) 'OTI-IER REFERENCES applica i Sobotka,Chemistry ofthe Sterids, 1938, page

